ObjectiveTo investigate the effect and potential mechanism of Zhishi Daozhi Formula （枳实导滞方， ZDF） on inflammation in rats with sepsis-induced gastrointestinal dysfunction based on the Toll-like receptor 4/myeloid differentiation factor 88/nuclear factor-κB （TLR4/MyD88/NF‑κB） signaling pathway and intestinal flora. MethodsSixty SD rats were randomly divided into sham‑operation group， model group， ulinastatin group， and low‑， medium‑ and high‑dose ZDF groups， with 10 rats in each group. Except for the sham‑operation group， the rat model of sepsis‑induced gastrointestinal dysfunction was established by cecal ligation and puncture （CLP）. Three days before modeling and 12 h after modeling， the low-， medium- and high-dose ZDF groups were intragastrically administered with ZDF solution at doses of 3.80， 7.60 and 15.20 g·kg⁻¹·d⁻¹， respectively. The ulinastatin group received intraperitoneal injection of ulinastatin at 100,000 U/kg， while the sham-operation and model groups were given normal saline at 1 ml/100 g by gavage. All interventions were performed once daily for 4 consecutive days. Tow hours after the last intervention， gastric emptying rate and small intestinal propulsion rate were detected. Gastrointestinal function indicators including D‑lactic acid （D‑LA） and intestinal fatty acid binding protein （i‑FABP）， as well as inflammatory indicators including procalcitonin （PCT） and C‑reactive protein （CRP） were measured by ELISA 24 h after modeling. The counts of Escherichia coli， Bifidobacterium and Lactobacillus in the intestine were compared. The histopathological morphological changes of the rat intestinal mucosa were observed by HE staining. The protein and mRNA expressions of IκB kinase （Ikk）， TLR4， MyD88 and NF‑κB in ileal tissues were detected by western blot and qRT‑PCR， respectively. ResultsCompared to the sham-operation group， the model group showed decreased gastric emptying rate and small intestinal propulsion rate， increased intestinal Escherichia coli count， elevated serum D‑LA， i‑FABP， PCT and CRP levels， increased protein and mRNA expressions of Ikk， TLR4， MyD88 and NF-κB in ileal tissue， decreased counts of Lactobacillus and Bifidobacterium （P＜0.05）， accompanied by obvious pathological injury of ileal mucosa. Compared to the model group， all medication groups exhibited significant improvements in the above mentioned indicators and alleviated intestinal mucosal injury （P＜0.05）. Compared to the ulinastatin group， except for D-LA in the medium-dose ZDF group， the differences in the above indicators between the low- and medium-dose ZDF groups were statistically significant （P＜0.05）， while the differences between the high-dose ZDF group and the ulinastatin group were not statistically significant （P＞0.05）. With increasing dosage， the gastric emptying rate， small intestinal propulsion rate， Lactobacillus and Bifidobacterium counts in the rats of all three ZDF groups increased， while the levels of serum D-LA， PCT， CRP， and i-FABP， the protein levels of Ikk， TLR4， MyD88， and NF-κB in intestinal tissue， and the Escherichia coli count decreased （P＜0.05）. ConclusionZDF can ameliorate intestinal mucosal injury， maintain intestinal flora balance and inhibit inflammatory response in rats with sepsis‑induced gastrointestinal dysfunction， with the high-dose group showing the best efficacy， which may be achieved by down‑regulating the TLR4/MyD88/NF-κB signaling pathway.